Cryptococcus neoformans is the etiologic agent of cryptococcosis, one of the most serious fungal diseases worldwide. Although C. neoformans primarily affects patients with impaired immune systems (AIDS, cytotoxic chemotherapy, corticosteroid therapy, sarcoidosis), people with no known underlying immunodeficiencies are also affected.. Meningoencephalitis, the most common clinical manifestation of cryptococcosis, is 100% fatal if not treated. Very little C. neoformans genome sequence is available. Before a detailed analysis of C. neoformans' molecular pathogenesis can be undertaken, C. neoformans' genes must be identified, and the genes sequenced. At present, the cheapest and fastest method to identify an organism's genes, and concomitantly to sequence the genes, is whole genome shotgun sequencing. We propose to shotgun sequence the whole C. neoformans genome to a coverage of a 4-to-5-fold in three years and to assemble the data. To that end, random small insert genomic libraries will be constructed in an M13- based vector and sequenced. The random sequence reads will be assembled. What will be produced is a "working draft" (in the newly revised Human Genome Project sense) of the C. neoformans genome. The working draft will be a robust platform upon which many experiments can be designed, and data interpreted: e.g., molecular pathogenesis, drug discovery, and vaccine development. An important and integral part of this proposed C. neoformans Genome Project is the immediate release of all sequence data. As we have done for other large scale genome/chromosome sequencing projects, we will post raw sequence data overnight. Contigs in progress will be posted immediately. Larger, more secure contigs will be posted and submitted to GenBank (HTGS phase 1) as soon as possible. Thus, all scientists worldwide will have immediate and equal access to the sequence data.